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WO1996017686A1 - Separation magnetique de spheres a sensibilite magnetique - Google Patents

Separation magnetique de spheres a sensibilite magnetique Download PDF

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Publication number
WO1996017686A1
WO1996017686A1 PCT/US1995/015146 US9515146W WO9617686A1 WO 1996017686 A1 WO1996017686 A1 WO 1996017686A1 US 9515146 W US9515146 W US 9515146W WO 9617686 A1 WO9617686 A1 WO 9617686A1
Authority
WO
WIPO (PCT)
Prior art keywords
magnetically responsive
population
container
homogeneous
homogeneous population
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US1995/015146
Other languages
English (en)
Inventor
Douglas Charles Gilbert
Peter George Cable
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
RTX BBN Technologies Corp
Original Assignee
Bolt Beranek and Newman Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Bolt Beranek and Newman Inc filed Critical Bolt Beranek and Newman Inc
Priority to AU42861/96A priority Critical patent/AU4286196A/en
Publication of WO1996017686A1 publication Critical patent/WO1996017686A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C1/00—Magnetic separation
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C1/00—Magnetic separation
    • B03C1/005—Pretreatment specially adapted for magnetic separation
    • B03C1/01—Pretreatment specially adapted for magnetic separation by addition of magnetic adjuvants
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C1/00—Magnetic separation
    • B03C1/02—Magnetic separation acting directly on the substance being separated
    • B03C1/28—Magnetic plugs and dipsticks
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/02—Devices for withdrawing samples
    • G01N1/10—Devices for withdrawing samples in the liquid or fluent state
    • G01N1/18—Devices for withdrawing samples in the liquid or fluent state with provision for splitting samples into portions
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/0098—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T436/00—Chemistry: analytical and immunological testing
    • Y10T436/25—Chemistry: analytical and immunological testing including sample preparation
    • Y10T436/25375—Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]

Definitions

  • the present invention relates to magnetic separation, and more particularly to the magnetic separation of magnetically responsive spheres admixed in a fluid.
  • a common method for identifying the presence of organic or inorganic substances in fluids involves capturing the substances with specially treated magnetically responsive spheres that are designed to attach themselves to a substance of interest.
  • the nature of the spheres allows for magnetic management of the spheres in that such spheres are highly permeable with low magnetic retentivity.
  • the spheres are responsive to and can be manipulated by magnetic fields, yet when the magnetic fields are removed the particles retain no magnetic properties.
  • Dynal beads Such spheres, or magnetically responsive particles, are commercially available, among other sources, from Dynal Inc., 45 North Station Plaza, Great Neck, N.Y., in 2.8 and 4.5 micron diameter sizes (hereinafter referred to as "Dynal beads” or “beads”) .
  • Dynal beads have been used as magnetically responsive beads for isolating, collecting and assaying diagnostic ligates as described in U.S. Patent No. 5,158,871 (“the '871 patent”) .
  • a suitable ligand is bound to a sheath of an organic substance surrounding the metal oxide core of the magnetically 96/17686 PCIYUS95/15146
  • the suitable ligand is capable of binding with a ligate or target substance that is sought to be isolated and is diagnostic of a particular disease state.
  • the ligand/beads are admixed with a fluid containing the ligate sought, for a selected time period and in a manner sufficient to effect a strong attachment between the ligate and ligand/beads to form ligate/ligand/bead complexes.
  • a magnetic gathering or harvesting device is contemplated in the '871 patent, to attract and retain the complexes in a localized magnetic field for removal from the fluid.
  • the '871 patent suggests only application specific, manually manipulated harvesting devices each suitable for a particular application.
  • the non-homogeneous population of target substance bound and unbound spheres is drawn in bulk toward and concentrated in the vicinity of a controller magnet.
  • Manual processing for the separation of the spheres can take many hours and is labor intensive. Only after considerable manual processing can spheres be separated from different populations for microscopic slide preparation. The spheres can then be tested to determine the presence of target substances attached to the surfaces of the spheres. To enhance the capture process, large numbers of spheres are often used.
  • spheres of different sizes, each treated for a different target substance may be used. In these cases, it is necessary and desirable, yet difficult, time consuming, labor intensive and costly, to separate spheres from different populations into homogeneous populations for further analysis.
  • the present invention provides a method and apparatus for automated separation of magnetically responsive spheres into different populations by carefully manipulating forces of gravity, buoyancy, fluid friction, and magnetism.
  • distinctly homogeneous sphere populations are separated from a non-homogeneous population comprised of a plurality of intermixed homogeneous sphere populations, without significant manual processing, by: magnetically effecting distancing of an initial non-homogeneous sphere population; facilitating enhanced separation of the non-homogeneous population in accordance with rate of descent of spheres within homogeneous populations; providing magnetic acceleration of the separation process; and effecting - A - magnetic concentration of the separated homogeneous sphere populations.
  • features of the process and apparatus according to the invention include benefits obtained in the art of bio-sensing resulting from the automated condensing of homogeneous groups of spheres from a non-homogeneous sphere population. Such benefits include improved detection, classification and guantitation of target substances. Laboratory test times are reduced due to the elimination of the requirement to test all sphere populations. Sampling efficiency is improved due to the ability to test for multiple target substances using spheres of different sizes. The testing process can be more fully automated by counting the separated spheres with magnetic or imaging techniques.
  • the method and apparatus can be implemented as a laboratory test device, an automated laboratory system, a portable field unit, or an in-line continuous real ⁇ time monitor. Further features include: exceptionally high separation efficiency and exceptionally rapid separation to facilitate exceptionally rapid examination. Additionally, unbound beads effectively separated can be recycled/reused.
  • FIG. 1A is a diagrammatic representation of an apparatus for containing a fluid with a target substance and magnetically responsive beads for implementing the method of separating the magnetically responsive beads according to the invention
  • Fig. IB is the apparatus of Fig. 1A wherein a disbursed bead cloud is formed
  • Fig. 1C is the apparatus of Fig. 1A wherein the magnetically responsive beads are subject to separation in accordance with relative rates of descent;
  • Fig. 2 is a graph illustrating calculated relative rates of descent of illustrative bead bound pathogens in an illustrative fluid.
  • an illustrative apparatus for performing the method of separating magnetically responsive beads generally includes a cylinder or column 10 which contains a fluid 12.
  • the column 10 has a substantially vertical orientation so as to define a top portion 14 and a bottom portion 16 thereof.
  • the top portion 14 includes a control magnet 16 that facilitates control of a non-homogeneous mixture of magnetically responsive beads 18 when it is desirable to retain the beads 18 at the top portion 14 of the column 10.
  • the control magnet can be a permanent magnet or an electromagnet, so long as its magnetic properties can be removed as discussed hereinafter.
  • the top portion 14 also includes a duct or port 20 for introducing the beads 18 and/or fluid 12 into the column.
  • the bottom portion 16 of the column 10 is preferably separable from the top portion 14 to facilitate removal of captured particles. Separability also permits easy access and cleaning of the interior of the column 10.
  • a concentrator magnet 22 is disposed proximate to the bottom portion 16 for expediting descent of beads within the column as discussed hereinafter.
  • a plurality of turns 24 of an electrically conductive wire coil are wound around the column 10 constituting an electromagnetic field generator that further includes a current source 26.
  • the current - fe - source 26 selectively delivers current to the coil turns 24 to provide an initial magnetic pulse and to set up electromagnetic lines of flux in a direction substantially parallel to a direction of descent of beads within the column.
  • the electromagnetic coil turns 24 are wound around the top portion 14 of the column 10 so as not to interfere with the separability of the top and bottom portions of the column.
  • the relative dimensions of the column top and bottom portions may differ from the present illustrative embodiment and the coil turns may extend to part of the bottom portion or be entirely disposed thereon.
  • the electromagnetic coil can be implemented in a Helmholtz configuration, having an appropriate gap between coil portions to facilitate visual or other observation, and simplify implementation of the process while providing a constant magnetic field.
  • a large quantity of the magnetically responsive beads or spheres, having a suitable outer sheath of bonding agent for binding with a target substance, are admixed in a fluid for a sufficient time and in a sufficient manner to allow for the spheres to come into contact with the target substance. It should be appreciated that such admixing can be done in the column 10, or in a container other than the column whereupon the beads or complexes would be collected, transported and injected into the column for the separation process. Once the complexing phase is complete, i . e .
  • the target substance has been sufficiently exposed to the magnetically responsive spheres with associated bonding agent within the fluid to form sphere/bonding agent/target substance complexes, it is desirable to effect control of the magnetically responsive spheres for commencement of the separation process.
  • the magnetically responsive spheres are disposed within the fluid 12 in the column 10 and retained against a horizontal surface via the control magnet 16 until the separation process begins.
  • the spheres are all clustered in the vicinity of the magnet, there is a tendency for the spheres to adhere to each other due to non-magnetic forces of compression and surface tension. This is especially true of spheres with diameters of a few microns.
  • the spheres are "magnetically responsive" and exhibit minimal magnetic retentivity, if any, there is a tendency for the spheres to adhere to one another even after removal of the control magnet.
  • the very small nature of the beads and the viscosity of the fluid creates a situation wherein large amounts of turbulence in the column, as would be created by stirring, shaking or the like, prolongs agitation of the beads and slows the separation process. It is preferable to create a localized agitation without creating great amounts of turbulence so that the viscous forces of the fluid in motion are not so dominant as to slow or impede the separation process.
  • the spheres will move to form chains that align with the flux lines of the applied field.
  • the chains will maintain cross-field separation due to the repelling forces of like dipoles.
  • the magnetic field impulse the magnitude of which is a function of the composition, size and quantity of beads, is applied only long enough to generate motion within the spheres, and then released.
  • the magnitude of the magnetic field impulse is controlled by the diameter and number of turns of the coil, and the current therethrough.
  • the sphere motion and resultant localized fluid motion generated by the field impulse will eliminate the clusters, effecting an initial separation and creating a cloud of beads at the top portion of the column.
  • the initial electromagnetic impulse excites each of the spheres into separate free-fall descent paths. Note that this concept may be applied to negatively buoyant spheres that fall through a fluid column, or to positively buoyant spheres that rise through a fluid column, or to combinations of positively and negatively buoyant spheres within a fluid column.
  • Further separation of the spheres is a function of the relative rates of descent (or ascent) of the beads or spheres.
  • Free falling (or free rising) spheres in the fluid reach terminal velocity which can be computed as a function of sphere mass, sphere diameter and roughness, and fluid density and viscosity.
  • the beads With no magnetic forces (other than gravitational forces) acting on the beads, as illustrated in Fig. 1C, the beads begin to fall as a distributed cloud and aggregate into homogeneous populations in accordance with common characteristics and rate of descent or motion within the particular fluid.
  • a free-fall period of time is required to allow for vertical separation of homogeneous groups. Vertical separation is a function of descent rate.
  • homogeneous groups are such that spheres within each group will have a common descent rate, while different homogeneous groups have different descent rates.
  • Many target substances may exhibit neutral buoyancy in the fluid in the column and act to slow the rate of descent of the attached bead, thus facilitating separation.
  • Fig. 2 shows a set of curves that describe the descent rates of bead-bound pathogens in water as a function of sphere mass and roughness and fluid density and viscosity calculations.
  • the microbeads are - s - typically 2.8, 4.5, or 10.0 microns in diameter as discussed hereinbefore, and when a bead comes in contact with the pathogen or target substance the bead attaches to the organism.
  • the negatively buoyant magnetically responsive beads descend at rates that relate to their particular condition, the populations will separate according to descent rates.
  • the descent rates of micro spheres can be very slow (100 to 500 microns per second) , so it is desirable to accelerate the process.
  • the separation process according to the invention is enhanced by magnetic acceleration.
  • a vertical and constant magnetic field can be applied to the fluid column.
  • the spheres in each of the separated populations will chain up in vertical orientation.
  • a general observation regarding the present illustrative embodiment is that beads that are not attached to a target substance aggregate as dipole "spears" which tend to descend faster than bead/target substance aggregates 28. In this orientation, the rates of descent will be magnified by the number of spheres in each chain, amplifying the separation process.
  • Application of a permanent magnet at the bottom of the fluid container will further amplify the separation speed because the spheres closest to the magnet will be drawn with the strongest magnetic force.
  • a second permanent magnet with equal magnetic force relative to the first magnet will be placed at the top of the fluid container.
  • the magnetic force of the bottom magnet will draw the fastest descent spheres to the bottom of the fluid column, and the magnetic force of the top magnet will draw the slowest descent beads to the top of the fluid column.
  • the separation process can be completed in a matter of minutes. A primary determinant of the time required is the length of the column, which is preferably as short as practicable so as to expedite the process.
  • the upper and lower magnets control the separated populations.
  • the fluid can be drawn off from the column and the upper and lower portions of the column separated to provide access to the populations for further preparation and testing.
  • a bead can be selected to combine with the magnetically responsive bead/target substance complex so as to effect a desired rate of descent and assure a high degree of homogeneous grouping in the descending beads. In such a case, the separation, acceleration and concentration aspects of the process according to the invention are not substantially changed.
  • a single coil with a plurality of turns is implemented in the illustrative embodiment shown and described herein to provide both the initial magnetic impulse and the vertically oriented magnetic field
  • different magnetic mechanisms can be implemented to effect both the initial magnetic impulse and the vertically oriented magnetic field.
  • a vertically or horizontally oriented electromagnet could be used to effect the electromagnetic impulse.
  • a combination - I I - of vertical and horizontal with alternating electromagnetic impulses could be used for initial separation
  • a separate Helmholtz coil or other magnetic mechanism could be used to effect the vertically oriented magnetic field for separation acceleration.
  • magnetic concentration can be effected at the sides, as opposed to the top of the column, and such concentration may be effected on a plurality of populations using a plurality of concentrator magnets in excess of two.
  • the column 10 as illustrated includes a single port or duct 20 for introducing beads and/or a fluid into the column, it will be appreciated that other means for introducing things into the vessel can be implemented, such as a plurality of ducts or ports, or hoses, spouts, conduits or the like.

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  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

Procédé et appareil de séparation automatique de sphères (18) à sensibilité magnétique portées dans un fluide, dans différentes populations, par manipulation prudente des forces de gravité, de flottabilité, de frottement visqueux, et de magnétisme. On sépare des populations de sphères homogènes distinctes (26) d'une population non homogène (28) constituée d'une pluralité de populations (26) de sphères homogènes mélangées, sans intervention manuelle importante, en établissant une distance magnétique d'une population (28) de sphères non homogène initiale, en facilitant une meilleure séparation de la population non homogène (28) selon une vitesse de descente de sphères à l'intérieur de populations homogènes (26), en produisant une accélération magnétique (24) dans le processus de séparation, puis en procédant à une concentration magnétique (22) des populations (26) de sphères homogènes séparées. Le processus et l'appareil assurent une biodétection améliorée résultant de la condensation automatique de groupes homogènes de sphères (26) à partir d'une population (28) de sphères non homogènes.
PCT/US1995/015146 1994-12-05 1995-11-21 Separation magnetique de spheres a sensibilite magnetique Ceased WO1996017686A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU42861/96A AU4286196A (en) 1994-12-05 1995-11-21 Magnetic separation of magnetically responsive spheres

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US08/349,627 1994-12-05
US08/349,627 US5628407A (en) 1994-12-05 1994-12-05 Method and apparatus for separation of magnetically responsive spheres

Publications (1)

Publication Number Publication Date
WO1996017686A1 true WO1996017686A1 (fr) 1996-06-13

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US (1) US5628407A (fr)
AU (1) AU4286196A (fr)
WO (1) WO1996017686A1 (fr)

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EP2338595A1 (fr) * 2009-12-23 2011-06-29 Atonomics A/S Dispositif, procédé et système pour la détection quantitative de la présence de plusieurs analytes cibles

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US6884357B2 (en) 1995-02-21 2005-04-26 Iqbal Waheed Siddiqi Apparatus and method for processing magnetic particles
WO1996026011A1 (fr) * 1995-02-21 1996-08-29 Siddiqi Iqbal W Appareil et procede de melange et de separation a l'aide de particules magnetiques
US6616623B1 (en) * 1997-07-02 2003-09-09 Idializa Ltd. System for correction of a biological fluid
US20040157219A1 (en) * 2003-02-06 2004-08-12 Jianrong Lou Chemical treatment of biological samples for nucleic acid extraction and kits therefor
FR2863626B1 (fr) * 2003-12-15 2006-08-04 Commissariat Energie Atomique Procede et dispositif de division d'un echantillon biologique par effet magnetique
US20050239091A1 (en) * 2004-04-23 2005-10-27 Collis Matthew P Extraction of nucleic acids using small diameter magnetically-responsive particles
WO2006017427A1 (fr) * 2004-08-03 2006-02-16 Becton, Dickinson And Company Utilisation de matières magnétiques pour fractionner des échantillons
EP1774334B1 (fr) * 2004-08-03 2017-10-04 Becton, Dickinson and Company Utilisation d'un materiau magnetique permettant de proceder a l'isolement de composes et le fractionnement d'echantillons constitues de plusieurs parties
US7147108B2 (en) * 2004-10-29 2006-12-12 Hewlett-Packard Development Company, Lp. Method and apparatus for the separation and collection of particles
WO2008067103A2 (fr) * 2006-10-30 2008-06-05 Stc.Unm Particules magnétiquement sensibles et appareils pour mélanger celles-ci
ES2665280T3 (es) * 2007-06-29 2018-04-25 Becton, Dickinson And Company Métodos para la extracción y purificación de componentes de muestras biológicas
US8292084B2 (en) * 2009-10-28 2012-10-23 Magnetation, Inc. Magnetic separator
JP5507432B2 (ja) * 2010-12-14 2014-05-28 シスメックス株式会社 分析装置および分析方法
WO2012145658A1 (fr) 2011-04-20 2012-10-26 Magnetation, Inc. Dispositif de séparation de minerai de fer
US9797817B2 (en) 2012-05-03 2017-10-24 The Regents Of The University Of Michigan Multi-mode separation for target detection

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Publication number Priority date Publication date Assignee Title
EP2338595A1 (fr) * 2009-12-23 2011-06-29 Atonomics A/S Dispositif, procédé et système pour la détection quantitative de la présence de plusieurs analytes cibles
WO2011076860A1 (fr) * 2009-12-23 2011-06-30 Atonomics A/S Dispositif, procédé, et système de détection quantitative de la présence de multiples analytes cibles

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Publication number Publication date
AU4286196A (en) 1996-06-26
US5628407A (en) 1997-05-13

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